Abstract Regulated neurotransmitter release is the basis for neuronal communication. Neurotransmitter is stored in synaptic vesicles and released via fusion with the plasma membrane. Consequently, the dynamics that govern synaptic vesicle morphology and function are deterministic for synaptic function. Synaptic vesicle dynamics can be divided into three segments: fusion, retrieval, and restoration. This application proposes to characterize these segments by capturing synaptic vesicles at accurate times after synaptic vesicle fusion and visualizing their dynamic states in the electron microscope. This approach will be complemented with diverse labeling techniques and genetically encoded markers for electron microscopy.